Heterovesicular liposomes

Abstract

Disclosed are heterovesicular liposomes containing substances of different biologically active compositions each encapsulated in separate chambers of the liposomes, having defined size distribution, adjustable average size, adjustable internal chamber size and number, methods of making them, and treatment of patients with them.

What is claimed is: 
     
       1. A heterovesicular liposome comprising at least two substances of different biological activity each encapsulated in separate chambers of the liposome. 
     
     
       2. The heterovesicular liposome of claim 1 wherein the encapsulated substance comprises a halogenohydroacid. 
     
     
       3. The heterovesicular liposome of claim 2 wherein the halogenohydroacid is selected from the group consisting of hydrochloric acid, hydroiodic acid, hydrobromic acid, hydrofluoric acid, and combinations thereof. 
     
     
       4. The heterovesicular liposome of claim 2 wherein the encapsulated substance comprises a hydrochloride. 
     
     
       5. The heterovesicular liposome of claim 4 wherein the hydrochloride is selected from the group consisting of lysine hydrochloride, histidine hydrochloride, arginine hydrochloride, glucosamine hydrochloride, quanine hydrochloride, and combinations thereof. 
     
     
       6. The heterovesicular liposome of claim 1 wherein the encapsulation is in a buffered medium. 
     
     
       7. A heterovesicular lipid vesicle or liposome produced by: the (a) encapsulating a first substance in the presence of an organic solvent by forming a first water-in-oil emulsion from two immiscible components containing a first substance to be encapsulated;   (b) encapsulating a second substance in the presence of an organic solvent by forming a second water-in-oil emulsion from two immiscible components containing a second substance to be encapsulated;   (c) encapsulating the product of step (a) and the product of step (b) by dispersing a chimeric emulsion of the product of step (a) and the product of step (b) in a third immiscible aqueous component to form solvent spherules; and   (d) removing organic solvent from the spherules to form the heterovesicular liposome.   
     
     
       8. The heterovesicular liposome of claims 1 or 7 wherein the biologically active substances are selected from the group consisting of antiasthmatics, cardiac glycosides, antihypertensives, antiparasitics, nucelic acids, antiarrhythmics, antianginas, hormones, antidiabetics, antineoplastics, tranquilizers, steriods, antihistamines, sedatives, analgesics, antibiotics, immunomodulators, antifungals, vasopressors, antivirals, vaccines, monoclonal antibodies, herbicides, neurotransmitters, radionuclides, and radio contrasts. 
     
     
       9. The heterovesicular liposome of claim 1 together with a targeted delivery system. 
     
     
       10. The heterovesicular liposome of claim 9 wherein the liposome is anatomically targeted. 
     
     
       11. The heterovesicular liposome of claim 9 wherein the liposome is mechanistically targeted. 
     
     
       12. The heterovesicular liposome of claim 9 wherein the liposome is passively targeted. 
     
     
       13. The heterovesicular liposome of claim 9 wherein the liposome is actively targeted. 
     
     
       14. The heterovesicular liposome of claim 13 wherein the liposome is actively targeted by coupling with a moiety selected from the group consisting of an antibody, a sugar, a glycolipid, and a protein. 
     
     
       15. A process for producing a heterovesicular lipid vesicle or liposome comprising the steps of: (a) encapsulating a first substance in the presence of an organic solvent by forming a first water-in-oil emulsion from two immiscible components containing a first substance to be encapsulated;   (b) encapsulating a second substance in the presence of an organic solvent by forming a second water-in-oil emulsion from two immiscible components containing a second substance to be encapsulated;   (c) encapsulating the product of step (a) and the product of step (b) by dispersing a chimeric emulsion of the product of step (a) and the product of step (b) in a third immiscible aqueous component to form solvent spherules; and   (d) removing organic solvent from the spherules to form the heterovesicular liposome.   
     
     
       16. The process of claim 15, wherein the first substance is encapsulated by providing a first lipid component dissolved in one or more organic solvents and adding into the first lipid component an immiscible first aqueous component containing the first substance and the second substance is encapsulated by providing a second lipid component dissolved in one or more organic solvents and adding into the second lipid component an immiscible second aqueous component containing the second substance. 
     
     
       17. The process according to claim 16 wherein the first and second lipid components are a phospholipid or an admixture of several phospholipids. 
     
     
       18. The process according to claim 17 wherein the phospholipids are selected from the group consisting of phosphatidylcholine, cardiolipin, phosphatidylethanolamine, sphingomyelin, lysophosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, and phosphatidic acid. 
     
     
       19. The process according to claim 17 wherein the one or more of the lipid components contain a lipid with a net negative charge or charges. 
     
     
       20. The process according to claim 17 wherein at least one of the phospholipids are provided in admixture with cholesterol. 
     
     
       21. The process according to claim 17 wherein at least one of the phospholipids are provided in admixture with stearylamine. 
     
     
       22. The process according to claim 15 wherein three or more water-in-oil emulsions containing three or more immiscible aqueous components are combined to form the chimeric emulsion. 
     
     
       23. The process according to claim 16 wherein the first and second lipid components are identical. 
     
     
       24. The process according to claim 16 wherein at least one of the first and second substances is a lipophilic biologically active material. 
     
     
       25. The process according to claim 16 wherein at least one of the first and second lipid components is a neutral lipid. 
     
     
       26. The process according to claim 16 wherein the organic solvent is selected from the group consisting of ethers, hydrocarbons, halogenated hydrocarbons, halogenated ethers, esters, and combinations thereof. 
     
     
       27. The process according to claim 15, wherein at least one substance to be encapsulated is a halogenohydroacid. 
     
     
       28. The process according to claim 27, wherein the halogenohydroacid is selected from the group consisting of hydrochloric acid, hydroiodic acid, hydrobromic acid, hydrofluoric acid and combinations thereof. 
     
     
       29. The process according to claim 15, wherein at least one substance to be encapsulated is a hydrochloride. 
     
     
       30. The process according to claim 29 wherein the hydrochloride is selected from the group consisting of lysine hydrochloride, histidine hydrochloride, arginine hydrochloride, glucosamine hydrochloride, guanine hydrochloride, and combinations thereof. 
     
     
       31. The process according to claim 15 wherein hydrophilic biologically active materials are the substances to be encapsulated. 
     
     
       32. The process according to claim 31 wherein the hydrophilic biologically active materials are selected from the group consisting of interleukin-2, cytosine arabinoside, methotrexate, 5-fluorouracil, cisplatin, floxuridine, melphalan, mercaptopurine, thioguanine, thiotepa, vincristine, vinblastine, streptozocin, leuprolide, interferon, calcitonin, doxorubicin, daunorubicin, mitoxanthrone, amacrine, actinomycin, and bleomycin. 
     
     
       33. The process according to claim 15 wherein the emulsification of the two components is carried out using methods selected from the group consisting of mechanical agitation, ultrasonic energy, and nozzle atomization. 
     
     
       34. The process according to claim 15 wherein the average size and number of the aqueous chambers within the liposome are determined by the type, intensity, and/or duration of the energy used in the formation of the emulsions. 
     
     
       35. The process according to claim 15 wherein the third aqueous component is a buffered medium. 
     
     
       36. The process according to claim 15 wherein the third aqueous component contains a neutralizing agent selected either singly or in combination from the group consisting of free-base lysine and free-base histidine. 
     
     
       37. The process according to claim 35 wherein the third aqueous component is an aqueous solution containing solutes selected from the group consisting of carbohydrates and amino acids. 
     
     
       38. The process according to claim 35 wherein the third aqueous component is an aqueous solution containing solutes selected either singly or in combination from the group consisting of glucose, sucrose, lactose, free-base lysine, and free-base histidine. 
     
     
       39. The process according to claim 15 wherein the dispersion to form solvent spherules is carried out using methods selected from the group consisting of mechanical agitation, ultrasonic energy, and nozzle atomization. 
     
     
       40. The process according to claim 39 wherein the average size of the liposome is determined by the type, intensity, and duration of the energy used. 
     
     
       41. The process according to claim 15 wherein the solvent is removed by evaporation. 
     
     
       42. The process according to claim 41 wherein the evaporation of the organic solvent is provided by passing gas over the second aqueous component. 
     
     
       43. The process of claim 15 wherein the first and second substances to be encapsulated are selected from the group consisting of antiasthmatics, cardiac glycosides, antihypertensives, antiparasitics, nucleic acids and analogs, antiarrhythmics, antianginas, hormones, antidiabetics, antineoplastics, tranquilizers, steroids, antihistamines, sedatives, analgesics, antibiotics, immunomodulators, antifungals, vasopressors, antivirals, vaccines, monoclonal antibodies, herbicides, neurotransmitters, radionuclides, and radio contrasts. 
     
     
       44. A method for the treatment of a patient with at least two biologically active compounds comprising, administering said compounds to the patient each encapsulated in separate chambers in a heterovesicular liposome. 
     
     
       45. A method for the treatment of a patient with at least two biologically active compounds comprising, administering to the patient heterovesicular liposomes encapsulating the biologically active substances according to claims 1 or 14. 
     
     
       46. The process according to claim 25 wherein at least one of the first and second lipid components further comprises a substance selected from the group consisting of vegetable oils, animal fats, tocopherols, tocopherol esters, cholesteryl esters, and hydrocarbons.